Utilizing rice straw and sugarcane bagasse as low-cost feedstocks towards sustainable production of succinic acid.

Succinic acid (SA) has been produced from rice straw (RS) and sugarcane bagasse (SB) as low-cost feedstocks in this study through sequential peracetic acid (PA) and alkaline peroxide (AP) pretreatment assisted by ultrasound and pre-hydrolysis followed by simultaneous saccharification and fermentation (PSSF). The effect of yeast extract (YE) concentration, inoculum concentration, and biomass type on SA production was investigated. The results showed that SA production from RS and SB was significantly affected by the YE concentration. Final concentration and yield of SA produced were significantly increased along with the increasing of YE concentration. Moreover, inoculum concentration significantly affected the SA production from SB. Higher inoculum concentration led to higher SA production. On the other hand, SA production from RS was not significantly affected by the inoculum concentration. Using RS as the feedstock, the highest SA production was achieved on the medium containing 15 g/L YE with 5 % v/v inoculum, obtaining SA concentration and yield of 3.64 ± 0.1 g/L and 0.18 ± 0.05 g/g biomass, respectively. Meanwhile, the highest SA production from SB was acquired on the medium containing 10 g/L YE with 7.5 % v/v inoculum, resulting SA concentration and yield of 5.1 ± 0.1 g/L and 0.25 ± 0.05 g/g biomass, respectively. This study suggested that RS and SB are potential to be used as low-cost feedstocks for sustainable and environmentally friendly SA production through ultrasonic-assisted PA and AP pretreatment and PSSF.

In-vitro antioxidant, in-vivo anti-inflammatory, and acute toxicity study of Indonesian propolis capsule from Tetragonula sapiens.

Propolis is widely used as traditional medicine since ancient times. It was necessary to conduct the pre-clinical study because of its relevant curative properties. This study aimed to investigate in-vitro antioxidant, standardize quality parameters, study acute toxicity, and determine in-vivo anti-inflammatory. Three spectrophotometric methods were used to determine antioxidant activity. The standardization includes physical, chemical, and microbiological evaluation. Furthermore, an acute toxicity test was conducted using 20 female Sprague Dawley (SD) strain rats divided into 4 groups with different dose of propolis. The in vivo anti-inflammatory test was carried out using the carrageenan induction method on rats' soles. A total of 36 female SD rats were classified into 6 groups as follows, Group normal, negative control, diclofenac sodium, and three propolis groups (72; 144; and 288 mg/kg BW). The results demonstrated the IC50 values of the DPPH and ABTS scavenging activity 9.694 ppm and 2.213 ppm, respectively. The FRAP reducing power was 189.05 mg AaE/g. The physical appearance of propolis capsule was vegicaps as white - white, size 0, with light brown granule. Moreover, the content weight was 418.88 mg with a disintegration time of 7 min 53 s, while the water, flavonoid, and polyphenol contents were 9.07%, 1.59%, and 98.0821 mg GAE/g respectively. The content of heavy metal and microbial contamination were not detected. The acute toxicity results showed LD50 ≥ 5 g/kg BW, no toxicity symptoms, and no abnormalities in all rats. The anti-inflammatory inhibition percentage for groups III, IV, V, and VI was 11.86%, 6.53%, 7.81%, and 6.63% respectively, while the anti-inflammatory drugs effectiveness percentage compared to positive controls were 55.00%, 65.83%, and 55.83% respectively. Based on these results, it can be concluded that propolis capsules fulfilled the standardization requirements, and it is likely to be non-toxic, and effective as antioxidant and anti-inflammatory.

Isolation of cyanide-degrading bacteria and molecular characterization of its cyanide-degrading nitrilase.

Hydrogen cyanide is an industrially important chemical, and its annual production is more than 1.5 million tons. Because of its toxicity, the cyanide-containing effluents from industries have caused many environmental problems. Among various methods to treat the contaminated soils or water, the biological degradation is regarded to be promising. We isolated two cyanide-degrading microorganisms, Pedobacter sp. EBE-1 and Bacillus sp. EBE-2, from soil contaminated with cyanide. Among these bacteria, Bacillus sp. EBE-2 exhibited significantly a high cyanide-degrading ability. Bacillus sp. EBE-2 might be used for the remediation of cyanide contaminated water or soil. A nitrilase gene was cloned from Bacillus sp. EBE-2. Bacillus nitrilase was expressed in Escherichia coli and purified. Bacillus nitrilase exhibited cyanide-degrading activity as a large oligomer. Since formic acid formation from cyanide was observed, Bacillus nitrilase is likely to be a cyanide hydrolase. Although there exist various homologous enzymes annotated as carbon-nitrogen family hydrolases, this is the first report on the cyanide degrading activity. The structure and catalytic site of Bacillus nitrilase were studied by homology modeling and molecular docking simulation.

Molecular docking and dynamics studies on propolis sulabiroin-A as a potential inhibitor of SARS-CoV-2.

Molecular docking and dynamics simulations were conducted to investigate the antiviral activity of Propolis Sulabiroin-A to inhibit the SARS-CoV-2 virus with quercetin, hesperidin, and remdesivir as control ligands. The parameters calculated were docking score and binding energy/molecular mechanics-generalized born surface area (MMGBSA), root mean square displacement (RMSD), and root mean square fluctuation (RMSF). Docking and MMGBSA scores showed that all the ligands demonstrate an excellent candidate as an inhibitor, and the order of both scores is hesperidin, remdesivir, quercetin, and sulabiroin-A. The molecular dynamics simulation showed that all the ligands are good candidates as inhibitors. Although the fluctuation of Sulabiroin-A is relatively high, it has less protein-ligand interaction time than other ligands. Overall, there is still a good possibility that sulabiroin-A can be used as an alternative inhibitor if a new structure of receptor SARS-CoV-2 is used.

Anti-Breast Cancer Activity on MCF-7 Cells of Melittin from Indonesia's Apis cerana: An In Vitro Study.

OBJECTIVE: Breast cancer is the most common case of cancers. Apitheraphy has been traditionally used for abundance diseases. This study aims to evaluate and compare the anti-breast cancer activity of melittin from Indonesia's Apic cerana as a potential drug for treating breast cancer. METHODS: Apis cerana bee venom (BV) was collected from a bee farm in Cikurutung, Bandung using an electrical venom device. The BV was then purified using the ÄKTA Start system and HiTrap™ SP HP cation exchange chromatography column. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) was used to identify melittin based on its molecular mass and lowry's protein assay to measure melittin concentration. Melittin cytotoxicity was measured with brine shrimp lethality test (BSLT), while MCF-7 breast cancer cells MTT assay was used to measure its anti-breast cancer activity, based on inhinition rate. RESULTS: 95.432 µg/mL melittin is purified from 62.8 mg/L BV, using  cation exchange chromatography. Melittin in vitro analysis with MCF-7 MTT assay is used to determine anti-breast cancer activity in dose dependent manner. Furthermore, melttin BSLT result showed a LC50 16.67675 µg/mL. Therefore, the MTT assay  was conducted in 5, 10 and 15 µg/mL with MCF-7 inhibition values of 0.768 ± 0.014, 3.303 ± 0.011, and 35.714 ± 0.009 %, respectively. CONCLUSION: Indonesia's Apis cerana has the potential to be used as a therapeutic peptide for breast cancer treatment.

Potential hepatoprotective effects of flavonoids contained in propolis from South Sulawesi against chemotherapy agents.

The use of doxorubicin and epirubicin as chemotherapy agent causes side effects such as liver damage due to oxidative stress by reactive oxygen species (ROS) that cause increased of ALT and AST level as liver parameter. One source of natural antioxidants as ROS neutralizer comes from flavonoid that contain in propolis. Most researchers claim that flavonoid can be used to protect the liver. The aim of this study was to test the hepatoprotective effect of flavonoid in propolis from South Sulawesi against doxorubicin and epirubicin. The experiment included male Sprague dawley rats divided into nine groups. The rats received the microcapsule propolis or the quercetin orally for 15 days. The hepatotoxicity was promoted by injection epirubicin and doxorubicin (i.v.) with a cumulative dose of 9 mg/kg. In this study, total polyphenol and flavonoid tests of propolis have been carried out, there were 1.1% polyphenols and 2.7% flavonoids, the antioxidant activity tests showed IC50 value of 9849 ppm and LCMS/MS tests supported the presence of phenolic compounds in propolis from South Sulawesi. Liver parameter was measured and the results showed that the propolis 200 mg/kg group produced the lowest ALT and had potential protective effect against doxorubicin and epirubicin-induced hepatotoxicity.

IN-SILICO dynamic analysis of Sulawesi propolis as anti-endometriosis drug: Interaction study with TNF alpha receptor, NF-kB, estrogen receptor, progesterone receptor and prostaglandin receptor.

INTRODUCTION: Endometriosis is a disease that impacts around 10% of all women in reproductive age, with pelvic pain and infertility as its main clinical features. Current medical treatment targeting lowering estrogen activity has not shown sufficient result due its side effects and reproductive function suppression. Propolis has been widely studied, showing anti inflammation and pro-apoptosis property, that could potentially be used in the treatment of endometriosis. This study investigates the interaction between Sulawesi Propolis' active components and receptors and protein related to endometriosis pathogenesis. METHODS: Active components of Sulawesi Propolis were initially identified with their targeted protein receptors. Lipinski rules were used to screen potential components. The ligands and proteins were tested using Autodock program to predict the most active compound and possible binding sites between propolis and some target proteins associated with inflammatory and apoptotic activity in endometriosis models. Receptor modelling is then performed using Swiss-Model. RESULTS: These active components of Sulawesi Propolis showed a strong binding potential towards TNF- α, NF-kb, Estrogen-α, Estrogen-ß, progesterone B, PGE2 EP2 and EP3 subtype respectively: Sanggenon C, Sanggenon H, Epicryptoacetalide, Chrysin-7-O-ß-D-glucopyranodside, Irilone, Polydatin and Epicryptoacetalide. Compared to its negative ligand, Sulawesi Propolis displayed a stronger binding capacity to TNF-α, Estrogen-α, and Progesterone B receptors. CONCLUSION: Sulawesi Propolis has the ability to interact with receptors related to reproductive function, apoptotic reactions and inflammatory processes, a significant factor associated with the pathogenesis of endometriosis.

Metabolome analysis and chemical profiling of Indonesian royal jellies as the raw material for cosmetic and bio-supplement products.

Royal jellies (RJs) possess moisturizing, emulsifying, and stabilizing properties, and several pharmacological activities have also been found to be present, which make them an ideal component for cosmetic and skin care products. However, despite the abundant efficacies, there is a lack of studies that explore the chemical composition of RJ using metabolome analysis. Furthermore, an evaluation of the chemical composition of Indonesian RJs collected from different regions has yet to be carried out. Therefore, the main objective of this study was to identify any differences in the chemical composition of such RJs. Chemical profiling was also carried out to enable more targeted utilization based on the actual compositions. Chemical profiling is also important given the rich Indonesian biodiversity and the high dependence of the RJ compositions on the botanical source. In this research, ultra-performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry was used as part of an untargeted metabolomics approach. From the chemical profiling, >30 compounds were identified across four RJ samples. The major constituents of the samples were found to be oligosaccharides, fatty acids, and adenosine monophosphate derivatives. Meanwhile, sucrose and planteose were found to be highest in the samples from Banjarnegara and Kediri, whereas dimethyloctanoic acid was found to be unique to the sample from Banjarnegara. It was also discovered that the RJs from Demak and Tuban contained more organic fatty acids and oligosaccharides than the other samples. Although the sample from Demak demonstrated good potential for use in the cosmetic, skin care, and bio-supplement industries, the higher abundance of fatty acids and oligosaccharides in the sample from Tuban indicated that it is perhaps the most suitable RJ for use in this field.

Evaluating the potency of Sulawesi propolis compounds as ACE-2 inhibitors through molecular docking for COVID-19 drug discovery preliminary study.

Coronavirus disease (COVID-19) is a global pandemic caused by severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2). Up to date, there has been no specific cure to treat the disease. Indonesia is one of the countries that is still fighting to control virus transmission. Yet, at the same time, Indonesia has a rich biodiversity of natural medicinal products that potentially become an alternative cure. Thus, this study examined the potency of a natural medicinal product, Sulawesi propolis compounds produced by Tetragonula sapiens, inhibiting angiotensin-converting activity enzyme-2 (ACE-2), a receptor of SARS-CoV-2 in the human body. In this study, molecular docking was done to analyze the docking scores as the representation of binding affinity and the interaction profiles of propolis compounds toward ACE-2. The results illustrated that by considering the docking score and the presence of interaction with targeted sites, five compounds, namely glyasperin A, broussoflavonol F, sulabiroins A, (2S)-5,7-dihydroxy-4'-methoxy-8-prenylflavanone and isorhamnetin are potential to inhibit the binding of ACE-2 and SARS-CoV-2, with the docking score of -10.8, -9.9, -9.5, -9.3 and -9.2 kcal/mol respectively. The docking scores are considered to be more favorable compared to MLN-4760 as a potent inhibitor.

Molecular interaction analysis of Sulawesi propolis compounds with SARS-CoV-2 main protease as preliminary study for COVID-19 drug discovery.

Coronavirus disease 2019 (COVID-19), a respiratory disease caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), is a global health concern, as the World Health Organization declared this outbreak to be a global pandemic in March 2020. The need for an effective treatment is urgent because the development of an effective vaccine may take years given the complexity of the virus and its rapid mutation. One promising treatment target for COVID-19 is SARS-CoV-2 main protease. Thus, this study was aimed to examine whether Sulawesi propolis compounds produced by Tetragonula sapiens inhibit the enzymatic activity of SARS-CoV-2 main protease. In this study, molecular docking was performed to analyze the interaction profiles of propolis compounds with SARS-CoV-2 main protease. The results illustrated that two compounds, namely glyasperin A and broussoflavonol F, are potential drug candidates for COVID-19 based on their binding affinity of -7.8 kcal/mol and their ability to interact with His41 and Cys145 as catalytic sites. Both compounds also displayed favorable interaction profiles with SARS-CoV-2 main protease with binding similarities compared to inhibitor 13b as positive control 63% and 75% respectively.

Surface plasmon resonance analysis for detecting non-structural protein 1 of dengue virus in Indonesia.

Dengue is an acute febrile disease caused by dengue virus (DENV) that is transmitted by Aedes sp., which causes serious health conditions in many countries. Non-structural protein 1 (NS1) is a co-factor for the RNA replication of this virus, which represents a new strategy for the identification of dengue. Prompt and accurate laboratory diagnosis of this infection is required to assist in patient triage and management, as well as prevent the spread of this infection. In the present study, we tested the potential of surface plasmon resonance (SPR) as a diagnostic tool for dengue infections. NS1 antigen protein was used as an analyte that targets anti-NS1 antibodies, with their interaction resulting in a change in the refractive index. In comparison to currently available gold-standard detection methods [enzyme-linked immunosorbent assay (ELISA) and reverse transcription polymerase chain reaction (RT-PCR)], SPR showed a similar sensitivity but greater efficiency and simplicity in terms of infection detection. Out of 26 samples collected from patients with dengue in Indonesia, SPR was able to correctly identify all 16 positively infected individuals at a lower concentration and a shorter period of time compared to ELISA and RT-PCR. This study revealed that SPR is a promising tool for DENV detection and potentially other diseases as well.

The Effects of stingless bee (Tetragonula biroi) honey on streptozotocin-induced diabetes mellitus in rats.

Diabetes mellitus (DM) is a metabolic disease characterised by chronic hyperglycaemia with impaired carbohydrate, fat and protein metabolism caused by defects in insulin secretion or action. Based on our previous research, stingless bee honey (SLBH) from Tetragonula biroi and T. laeviceps can inhibit alpha-glucosidase activities. Therefore, the aim of the present study was to determine the effects of daily oral administration of SLBH on body weight (BW) and fasting blood glucose (FBG) levels of male rats with streptozotocin (STZ)-induced DM. Thirty-six male Sprague Dawley rats were divided into six groups of six rats each. One group of normal non-diabetic rats served as a positive control. The diabetic groups were intraperitoneally (i.p.) injected with STZ (50 mg/kg BW) for induction of DM and divided into five equal subgroups of six animals each: an untreated group as a negative control; a group treated with 0.6 mg/kg BW of glibenclamide as a positive control and three SLBN treatment groups that had daily oral administration of 0.5, 1.0 or 2.0 g/kg BW, respectively, for 35 days. The results showed that SLBH significantly reduced loss of BW in diabetic rats. FBG levels in diabetic rat blood, collected from the tail, were measured using Accu-Chek test strips. The FBG levels in diabetic rats that have oral administered intake with glibenclamide and SLBH were stable. There were no changes in serum FBG levels in SLBH-treated diabetic rats for 35 days. Pancreatic histopathology results from all groups showed no abnormalities or tissue damage in either diabetic or non-diabetic rats. The results of this study show that administration of SLBH reduced BW loss or improved BW of rats with STZ-induced DM. Meanwhile, the reduction in loss of BW that occurred in diabetic rats after 35 days of SLBH administration was the result of reduced formation of fats and proteins, which are broken down into energy. Further research is needed to determine the antidiabetic effects of honey from other stingless honeybee species.

Immobilization of Cholesterol Oxidase from Streptomyces Sp. on Magnetite Silicon Dioxide by Crosslinking Method for Cholesterol Oxidation.

Enzymatic biosensor has been paid much attention to the research fields due to its advantage in medical application. As one of the application, we determined the optimum value of cholesterol oxidase against cholesterol. In this work, we studied the behavior of cholesterol oxidation by enzymatic reaction to get the optimum condition for cholesterol oxidation. The enzyme that used were in two form, free cholesterol oxidase, and immobilized cholesterol oxidase. Cholesterol oxidase was produced from Streptomyces sp. by using solid state fermentation method and identified had high enzyme activity to be 5.12 U/mL. Cholesterol oxidase was simultaneously crosslinked immobilized onto magnetite coated by silicon dioxide (M-SiO2). The support was characterized by Fourier transform infrared (FTIR) to determine the functional group of modified particle and scanning electron microscope (SEM) to observe the morphological or our prepared particle. Cholesterol oxidase sensitivity to substrate was analyzed by using HPLC with different interval time measurements. The oxidation of cholesterol by free enzyme and immobilized enzyme was also investigated. The best sensitivity of cholesterol oxidase was estimated to oxidize Cso (concentration of substrate) 1.46 mM of substrate with Ce (concentration of enzyme) 20 mg/mL for 180 min. Final oxidation value of cholesterol by immobilized enzyme was greater than 60%. The results of this study revealed that immobilized enzyme for cholesterol oxidation was stable, reproducible, and sensitive.

The beneficial effect of Indonesian propolis wax from Tetragonula sp. as a therapy in limited vaginal candidiasis patients.

Vaginal candidiasis characterized by abnormal vaginal discharge and itching usually treated by azole's drug or nystatin; however, some results of treatment are unsatisfied and become recurrent. Propolis containing polyphenols and flavonoids is known to have anti-inflammatory and antimicrobial activity. This study investigated the effect of Indonesian propolis wax from Tetragonula sp. as a therapy in limited vaginal candidiasis patients. The subjects were women who came to the Tasik Community Health Centre met the inclusion criteria such as clinical complaint and laboratory evaluation (positive hyphae/pseudohyphae and culture on Sabouraud Dextrose Agar (SDA) medium) from a vaginal swab. Evaluation of anti-candida effect of propolis was determined by clinical remission and the absence of Candida's growth on SDA medium. Forty subjects were randomly assigned to those receiving treatment by ovule propolis (n = 20) and that treatment by nystatin (n = 20) as a control, once daily, for seven days, respectively. All methods have been approved by the Ethics Commission of the Faculty of Medicine, Universitas Indonesia. Our results indicated no significant difference in the laboratory evaluation of patients who have treated ovule propolis compared to standard therapy. This study suggests that propolis wax has a beneficial effect to develop as an anti-candida agent for vaginal candidiasis therapy.

Anti-inflammatory activity of Tetragronula species from Indonesia.

Anti-inflammatory drugs inhibit inflammation, particularly those classified as nonsteroidal anti-inflammatory drugs (NSAIDs). Several studies have reported that propolis has both anti-ulcerogenic and anti-inflammatory effects. In this study, we investigated the bioactive compound and in vivo anti-inflammatory properties of both smooth and rough propolis from Tetragronula sp. To further identify anti-inflammatory markers in propolis, LC-MS/MS was used, and results were analyzed by Mass Lynx 4.1. Rough and smooth propolis of Tetragonula sp. were microcapsulated with maltodextrin and arabic gum. Propolis microcapsules at dose 25-200 mg/kg were applied for carrageenan-induced rat's paw-inflammation model. Data were analyzed by one-way ANOVA and Kruskal-Wallis statistical tests. LC-MS/MS experiments identified seven anti-inflammatory compounds, including [6]-dehydrogingerdione, alpha-tocopherol succinate, adhyperforin, 6-epiangustifolin, deoxypodophyllotoxin, kurarinone, and xanthoxyletin. Our results indicated that smooth propolis at 50 mg/kg inhibited inflammation to the greatest extent, followed by rough propolis at a dose of 25 mg/kg. SPM and RPM with the dose of 25 mg/kg had inflammatory inhibition value of 62.24% and 58.12%, respectively, which is comparable with the value 70.26% of sodium diclofenac with the dose of 135 mg/kg. This study suggests that propolis has the potential candidate to develop as a non-steroid anti-inflammatory drug.

Purification and characterization of proteins in multifloral honey from kelulut bee (stingless bee).

The kelulut bee (Meliponini) is a subfamily of stingless bees that produce honey. A total of 89 species out of a total of 500 species of kelulut bees are known to originate from the Indo-Australian region. Kelulut bees do not have quality standards so they still refer to the Codex and EU Directive which basically only applied for Apis honey. The Codex and EU Directive are formed by several psychochemical parameters, one of it is diastase activity. Diastase activity in kelulut honey is known not to meet existing standards or even undetectable. Therefore, this study aimed to explore proteins inside kelulut honey and investigate the possibility of using a specific protein as a biomarker to differentiate honey produced by kelulut bee from other honey. This research can also be considered as an initial step to optimize the exploration of protein in kelulut honey. This research is divided into two sections which are the preliminary research and the research expansion. From preliminary section, glucose dehydrogenase enzyme (GDH) was found to be present inside Tetragonula spp honey. A further examination of GDH enzyme was made in four kelulut bee honeys namely Tetragonula leaviceps, T. biroi, Heterotrigona itama, and Geniotrigona thoracica. The preliminary research has five stages that are exactly the as expansion research section except it didn't include GDH activity measurement. The research includes seven main stages. First honeys were dialyzed to remove the sugar content followed by centrifugation. The samples were then purified using liquid chromatography with anion exchanger column. The molecular weight of proteins was analysed by SDS-PAGE method. The GDH activity was measured using spectrophotometer followed by qualitative analysis using LC-MS/MS. The peptide sequences resulted from LC-MS/MS were then matched with Uniprot to identify the unknow protein. The results showed that only T. biroi and T. laeviceps had GDH enzyme activity of 0,1891 U/mL and 0,1652-1,579 U/mL, respectively. Bands from both species were also qualitatively identified as GDH. With these results, it can be concluded that the GDH enzyme cannot be used as a biomarker to distinguish the kelulut honey.

Identification and classification of honey's authenticity by attenuated total reflectance Fourier-transform infrared spectroscopy and chemometric method.

BACKGROUND AND AIM: The authentication of honey is important to protect industry and consumers from such adulterated honey. However, until now, there has been no guarantee of honey's authenticity, especially in Indonesia. The classification of honey is based on the bee species (spp.) that produces it. The study used honey from sting bee Apis spp. and stingless bee Tetragonula spp. based on the fact that the content off honey produced between them has differences. Authenticating honey with currently available rapid detection methods, such as 13C nuclear magnetic resonance analysis, is costly. This study aimed to develop an inexpensive, fast, precise, and accurate classification method for authenticating honey. MATERIALS AND METHODS: In this study, we use attenuated total reflectance Fourier-transform infrared (ATR-FTIR) spectroscopy with wavelengths ranging between 550 and 4000 cm-1 as an alternative analysis method, which is relatively less expensive. The spectra of authentic and fake honey samples were obtained using ATR-FTIR and plotted using chemometric discriminant analysis. The authentic honey samples were acquired from a local Indonesian breeder of honey bees, while the fake honey samples were made from a mixture of water, sugar, sodium bicarbonate, and authentic honey. Data were collected using Thermo Scientific's OMNIC FTIR software and processed using Thermo Scientific's TQ Analyst software. RESULTS: Our method effectively classified the honey as authentic or fraudulent based on the FTIR spectra. To authenticate the honey, we formed two classes: Real honey and fake honey. The wavelengths that can best differentiate between these two classes correspond to four regions: 1600-1700 cm-1; 1175-1540 cm-1; 940-1175 cm-1; and 700-940 cm-1. Similarly, for classification purpose, we formed two classes: Apis spp. and Tetragonula spp. The wavelength region that can best classify the samples as belonging to the Apis spp. or Tetragonula spp. class is explicitly within the range of 1600-1700 cm-1. CONCLUSION: This study successfully demonstrated a method to rapidly and accurately classify and authenticate honey. ATR-FTIR is a useful tool to test the authenticity of honey.

Preliminary studies: the potential anti-angiogenic activities of two Sulawesi Island (Indonesia) propolis and their chemical characterization.

Several studies have previously reported propolis, or its constituents, to inhibit tumour angiogenesis. The anti-angiogenic activity of two Indonesian stingless bee propolis extracts from Sulawesi Island on vascular cells were assessed. Sample D01 was obtained from the outer side of bee hives, while D02 was from the inner side of the same hives. The extracts were profiled by using liquid chromatography coupled to high resolution mass spectrometry. The anti-angiogenic capacity was assessed on HUVECs and placenta-derived pericytes by cell viability, multi-channel wound healing, and CoCl2 based-hypoxia assays. The exact chemical composition has not been confirmed. The most abundant compounds in Indonesian sample D01 seem to be unusual since they do not immediately fall into a clear class. Two of the most abundant compounds have elemental compositions matching actinopyrones. Identification on the basis of elemental composition is not definitive but compounds in D01 are possibly due to unusually modified terpenoids. Sample D02 has abundant compounds which include four related diterpenes with differing degrees of oxygenation and some sesquiterpenes. However, again the profile is unusual. The anti-angiogenic assays demonstrated that D01 elicited a strong cytotoxic effect and a considerable anti-migratory activity on the vascular cells. Although D02 demonstrated a much weaker cytotoxic effect on the cell lines compared to D01, it elicited a substantial protective effect on the pericytes against CoCl2-induced dropout in an experiment to mimic a micro-environment commonly associated with angiogenesis and tumour growth. These results demonstrate modulatory effects of these propolis samples in vascular cells, which requires further investigation.

Antiretroviral activity of Pterois volitans (red lionfish) venom in the early development of human immunodeficiency virus/acquired immunodeficiency syndrome antiretroviral alternative source.

AIM: This study aimed to investigate the antiviral activity of Pterois volitans phospholipase A2 (PV-PLA2) from Indonesia to human immunodeficiency virus (HIV). MATERIALS AND METHODS: Fresh venomous fin parts of wild PV specimens were collected from Java Sea waters. Then, it washed using phosphate buffer pH 7.0 and immersed in phosphate buffer pH 7.0 0.01 m containing CaCl2 0.001 m for 24 h. The immersed fin then allowed for extraction process by sonicating for 2×8 min with 80% pulse and 20 kHz output with temperature controlling to avoid denaturation. The crude venom (CV) extracted from the fin is allowed for purification by 80% ethanol (ET) precipitation and ammonium sulfate fractionation method. The purified PV-PLA2 then analyzed using Lowry's method, Marinette's method, sodium dodecyl sulfate-polyacrylamide gel electrophoresis, and 3-(4, 5-dimethyl thiazol-2yl)-2, 5-diphenyl tetrazolium bromide assay. After determining the purest and safest sample of six samples analyzed, the chosen sample then tested into simian retrovirus-2 (SRV2)-A549 culture (48×104 cells/mL at 1-4 ppm), and compared to the CV sample (1-4 ppm) and lamivudine (100 ppm). The culture then is analyzed using a quantitative real time-polymerase chain reaction to find out the copy number of SRV-2 virus in each culture. RESULTS: The protein's activity, concentration, and purity analysis revealed that the PV-PLA2 purified using ammonium sulfate fractionation has the highest activity (1.81 times higher than the CV at 80% fractionation) and has higher purity than the sample from ET fractionation. The testing of the sample purified using ammonium sulfate fractionation at 80% saturation level shown that it has a 97.78% inhibition level toward SRV2-A549 culture at 4 ppm. However, in comparison to lamivudine which has 99.55% inhibition level at 100 ppm, it needs much lower concentration to achieve the same result. CONCLUSION: The significant inhibition of SRV2-A549 culture shown that the PV-PLA2 extracted from PV venom has the potential to become anti-HIV substances. It would be worthwhile to further evaluate the antiretroviral activity of PV-PLA2 in the in vivo studies.